RNA干扰GCF2基因沉默对人肝癌BEL-7404细胞周期的影响

发布时间：2018-03-05 01:28

本文选题：GC结合因子　切入点：肝细胞癌　出处：《医学研究生学报》2017年03期 　论文类型：期刊论文

【摘要】：目的 GC结合因子2(GCF2)是一种转录抑制因子,通过特异性结合富含GC序列的基因启动子而抑制该基因转录。文中通过靶向沉默GCF2基因表达观察GCF2对人肝癌细胞BEL-7404细胞周期的影响。方法将细胞分为空白对照组、阴性对照组及GCF2-siRNA组。空白对照组不转染任何序列,阴性对照组转染阴性对照序列,GCF2-siRNA组转染靶向GCF2基因的GCF2-siRNA序列以沉默GCF2在肝癌BEL-7404细胞中的表达。CCK-8法和流式细胞术分别检测沉默GCF2对细胞增殖和周期的影响,Western blot检测细胞周期蛋白cyclin D1及cyclin E、细胞周期蛋白依赖性激酶Cdk2及Cdk4蛋白表达。结果 GCF2下调后,GCF2-siRNA组细胞增殖在转染24、48、72和96 h受到不同程度抑制,抑制率分别为27.3%,43.6%,33.5%和29.7%,以转染48 h抑制率最高。与GCF2-siRNA组G1期百分比[(61.50±0.47)%]比较,空白对照组、阴性对照组[(45.50±0.75)%、(47.30±2.44)%]均下降(P0.05);与GCF2-siRNA组S期百分比[(21.30±1.23)%]比较,空白对照组、阴性对照组[(37.00±1.80)%、(34.50±1.86)%]均升高(P0.05)。转染48 h后,GCF2-siRNA组cyclin E和Cdk2蛋白相对表达量明显低于阴性对照组和空白对照组(P0.05)。结论下调GCF2表达引起BEL-7404细胞周期阻滞,提示GCF2对肝癌细胞周期影响可能是其参与肝癌发生发展的重要途径。
[Abstract]:Objective GC binding factor 2 (GCF2) is a transcription suppressor. The transcription of the gene was inhibited by binding to a gene promoter rich in GC sequence. The effect of GCF2 on the cell cycle of human hepatocellular carcinoma cell line BEL-7404 was observed by silencing the expression of GCF2 gene. Methods the cells were divided into blank control group. Negative control group and GCF2-siRNA group. Blank control group does not transfect any sequence, Transfection of GCF2-siRNA sequence targeting GCF2 gene in negative control group by silencing GCF2 expression in BEL-7404 cells. CCK-8 method and flow cytometry were used to detect the effect of silencing GCF2 on cell proliferation and cell cycle. Western blot was used to detect the effect of silencing GCF2 on cell proliferation and cell cycle. Results the expression of cyclin D1 and cyclin E, cyclin dependent kinase Cdk2 and Cdk4 protein in GCF2-siRNA group after GCF2 down-regulation was inhibited in different degree at 72 and 96 h after transfection with GCF2-siRNA. The inhibition rates were 27.33% and 29.7%, respectively. The inhibition rate was the highest in 48 hours after transfection. Compared with the GCF2-siRNA group, the percentage of G1 phase [61.50 卤0.47%] in the blank control group, the negative control group [45.50 卤0.75 卤47.30 卤2.44%] decreased P0.05%, compared with the S phase percentage of GCF2-siRNA group [21.30 卤1.23], the blank control group. The relative expression of cyclin E and Cdk2 protein in GCF2-siRNA group was significantly lower than that in negative control group and blank control group 48 h after transfection. Conclusion down-regulation of GCF2 expression leads to BEL-7404 cell cycle arrest. The results suggest that the effect of GCF2 on the cell cycle of HCC may be an important way to participate in the development of HCC.
【作者单位】：桂林医学院组织胚胎学教研室;桂林医学院临床医学院;
【基金】：广西壮族自治区高校科学技术研究项目(KY2015LX284) 大学生创新创业训练计划项目(201510601003)
【分类号】：R735.7

【相似文献】